CN108217972A - A kind of application of immobilization laccase in gold orange II is decolourized - Google Patents
A kind of application of immobilization laccase in gold orange II is decolourized Download PDFInfo
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- CN108217972A CN108217972A CN201711390178.1A CN201711390178A CN108217972A CN 108217972 A CN108217972 A CN 108217972A CN 201711390178 A CN201711390178 A CN 201711390178A CN 108217972 A CN108217972 A CN 108217972A
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- carboxyl carbon
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/342—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the enzymes used
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/0004—Oxidoreductases (1.)
- C12N9/0055—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10)
- C12N9/0057—Oxidoreductases (1.) acting on diphenols and related substances as donors (1.10) with oxygen as acceptor (1.10.3)
- C12N9/0061—Laccase (1.10.3.2)
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y110/00—Oxidoreductases acting on diphenols and related substances as donors (1.10)
- C12Y110/03—Oxidoreductases acting on diphenols and related substances as donors (1.10) with an oxygen as acceptor (1.10.3)
- C12Y110/03002—Laccase (1.10.3.2)
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/308—Dyes; Colorants; Fluorescent agents
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- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
A kind of application the invention discloses immobilization laccase and its in water process, the immobilization laccase is using the composite material prepared by polymethyl methacrylate is coated on carboxyl carbon nano tube surface as carrier, glutaraldehyde is bonding agent, and laccase is fixed in the carboxyl carbon nanotube coated with polymethyl methacrylate using covalent coupling method.The carboxyl carbon nanotube fixing laccase that the immobilization laccase of the present invention is coated based on porous material polymethyl methacrylate, so that the load factor height of laccase, can play absorption and the effect of catalytic degradation simultaneously has many advantages, such as that physical and chemical performance is stable to the progress efficient decolorizing processing of gold orange II, carries enzyme rate height, high recycling rate.
Description
Technical field
The present invention relates to a kind of application of immobilization laccase in gold orange II is decolourized, and belong to the water process neck in environmental protection
Domain.
Background technology
Laccase is a kind of polyphenol oxidase of cupric, by peptide chain, sugared aglucon and Cu2+3 parts form.Laccase is in nature
It is distributed widely in boundary in plant, fungi, a small number of insects and bacterium, laccase is had also discovered in some animal kidneys and serum.
Laccase can be catalyzed the oxidation reaction of many compounds, and substrate specificity is than wide, including amino-phenol, polyphenol, polyamines, wooden
Element, aryl diamine, vanillic acid, veratryl alcohol, ferulic acid and tween etc., and reaction product is water, green, environmental protection.Therefore, laccase quilt
It is widely used in various fields, such as：Paper pulp papermaking field, biosynthesis field, field of food, energy field, timber processing
Field, improves fiber properties field, field of biological detection etc. at field of environment protection.But laccase is easily inactivated by such environmental effects,
The shortcomings of not reproducible utilization.
Enzyme immobilizatio is to fetter enzyme with carrier material or be limited in certain area, can still provide for its distinctive catalysis
Reaction, and can realize that enzyme repeats continuous use and improved stability one kind technology.Process for fixation include adsorption method,
Investment, covalent coupling method and system blocking cut-off process.Adsorption method is to utilize the natural or artificial conjunction with many aperture structures
It is attached on the carrier with dissimilar charges into material adsorptive enzyme to carrier surface or by enzyme by electrostatic attraction and is allowed to solid
It is fixed.Investment be by cell encapsulation in gel network structure or semipermeable polymeric film or make cellular invasion enter porosity load
Internal portion.Covalent coupling method, which refers to, in the characteristic or function using special reagent, makes the reactive group on enzyme surface directly be sent out with reagent
Raw reaction, the immobilised enzymes of reticular structure is formed by being cross-linked with each other for group.System blocking cut-off process refers to using various films, such as
Cell is limited to the method in specific spatial dimension by the semi-permeable membranes such as dialyser, ultrafiltration membrane, hollow-fibre membrane.
Waste water from dyestuff largely enters China's water body environment, it has also become one of an important factor for polluting China's quality of water environment.
There are technical difficulties for the mineralising of removing and complicated hardly degraded organic substance of the outstanding problem of dying industrial wastewater processing including coloration.
Current most widely used dyestuff can be divided into three classes：Azo dyes, anthraquinone dye and triphenylmethane dye.In order to realize
The decoloration of waste water from dyestuff and mineralising efficient process need to start with from the microstructure of dyestuff, its degradation mechanism are analyzed, and
Develop the preferable dye wastewater treatment technology of specific aim.Gold orange II belongs to azo dyes, while is also anionic dye.
Invention content
It is an object of the invention to study the immobilization laccase of carboxyl carbon nanotube coated with polymethyl methacrylate
Decolorizing effect for anionic dye gold orange II and the application in wastewater treatment.
The present invention provides a kind of application of immobilization laccase in gold orange II is decolourized, and includes the following steps：
（1）It weighs the above-mentioned carboxyl carbon nanotubes obtained by the reaction of 0.2 g to be distributed in the normal octane solution of 5 mL 6%, adds in 0.2
G lauryl sodium sulfate and 30 mL water, molten ultrasonic 15 min of ice；
（2）4 mL methyl methacrylates, 1 mL divinylbenzenes, 8 mL1% potassium peroxydisulfates are added in, 70 DEG C of reaction temperature is stirred
24 h of polymerization are mixed, after the completion of reaction, are naturally cooling to room temperature, are centrifuged；
（3）The obtained solid water and ethyl alcohol of above-mentioned reaction alternately wash, and are dried in vacuo, cooling, collect product and obtain black
Solid particle is the carboxyl carbon nanotube coated with polymethyl methacrylate；
（4）The carboxyl carbon nanotube that 0.2 g polymethyl methacrylates coat is weighed, adds in the glutaraldehyde water of 20 ml 10%
Solution, room temperature mechanical stir 6 h, after the completion of reaction, are washed with water；
（5）The material of synthesis is added in 30 ml distilled water, adds in 0.02 g of laccase powder, it is 120 to be then placed in rotating speed
In the shaking table of rpm, 30 DEG C of 6 h of reaction；
（6）Finally with PH=7.0, three times, removal fails to combine resolvase up, receives for the phosphate buffer washing of 0.1 mol/l
Collect immobilization laccase, it is spare.
（7）0.05 g immobilization laccases are weighed, is put into II solution of gold orange of a concentration of 10 mg/L of 20 mL, is in rotating speed
120 r/min, temperature are 30 DEG C, and 24 h are reacted in pH=7.Then it collects supernatant and surveys absorbance.
Step（1）Described in ultrasound be first ultrasound 2 min when normal octane solution is distributed to carboxyl carbon nanotube, guarantee
Then the dispersibility of carboxyl carbon nanotube in the solution adds other reagents and carries out molten ultrasonic 15 min of ice.
Step（1）Used in carboxyl carbon nanotube have larger specific surface area, polymethyl methacrylate can be made
In sufficient dispersin polymerization to carboxyl carbon nanotube, the surface area of carrier is improved.
Step（1）In the dispersion liquid of prepared carboxyl carbon nanotube in 6% normal octane a concentration of 40 mg/ml.
Step（2）Middle polymeric reaction temperature should be always at 70 DEG C.
Step（3）It is middle that 6 h first need to be fully impregnated with ethyl alcohol in water and ethanol wash process, ensure fully removal impurity.
Step（5）The mass ratio of middle polymethyl methacrylate and laccase is 10：1.
Step（7）Middle supernatant absorbance of surveying is the concentration in order to obtain II solution of the gold orange after decolorization, and then calculate
Go out percent of decolourization.
This immobilization laccase prepared by the present invention is characterized in that：The carboxyl carbon nanometer of polymethyl methacrylate cladding
Pipe plays the role of adsorbent in system, can be conducive to methylene blue in carrier with absorbed portion laccase, porous structure
The enrichment on surface.Polymethyl methacrylate is coated on carboxyl carbon nanotube, can be effectively increased surface area, is glutaraldehyde and paint
Combination between enzyme provides a large amount of reaction site.
The advantage of the invention is that：Carboxyl carbon nanotube is coated on polymethyl methacrylate, wherein carboxylic can be played
The advantage of base carbon nanotube large specific surface area can adsorb laccase, increase and carry enzyme rate, be enriched with more gold oranges II, and can pass through
Having coated polymethyl methacrylate increases surface area, and more multiple binding sites are provided for glutaraldehyde and laccase, has physical chemistry
Performance is stable, carry enzyme rate height, high recycling rate, handles the features such as efficient.
Description of the drawings
Fig. 1 is the scanning electricity for the carboxyl carbon nanotube coated with polymethyl methacrylate that the embodiment of the present invention 1 provides
Mirror figure；
Fig. 2 is the X-ray diffractogram of carboxyl carbon nanotube coated with polymethyl methacrylate that the embodiment of the present invention 2 provides
Spectrum；
Fig. 3 is the infrared spectrum of carboxyl carbon nanotube coated with polymethyl methacrylate that the embodiment of the present invention 3 provides；
Fig. 4 (a) is the percent of decolourization that the immobilization laccase that the embodiment of the present invention 4 provides varies with temperature gold orange II；
Fig. 4 (b) is that the immobilization laccase that the embodiment of the present invention 4 provides changes gold orange II with the increased percent of decolourization of access times.
Specific embodiment
Above-mentioned purpose, feature and advantage to enable invention are more obvious understandable, below to the specific implementation of the present invention
Mode is described in detail.
Embodiment 1：
（1）It weighs the above-mentioned carboxyl carbon nanotubes obtained by the reaction of 0.2 g to be distributed in the normal octane solution of 5 mL 6%, adds in 0.2
G lauryl sodium sulfate and 30 mL water, molten ultrasonic 15 min of ice；
（2）4 mL methyl methacrylates, 1 mL divinylbenzenes, 8 mL1% potassium peroxydisulfates are added in, 70 DEG C of reaction temperature is stirred
24 h of polymerization are mixed, after the completion of reaction, are naturally cooling to room temperature, are centrifuged；
（3）The obtained solid water and ethyl alcohol of above-mentioned reaction alternately wash, and are dried in vacuo, cooling, collect product and obtain black
Solid particle is the carboxyl carbon nanotube coated with polymethyl methacrylate；
（4）The carboxyl carbon nanotube that 0.2 g polymethyl methacrylates coat is weighed, adds in the glutaraldehyde water of 20 ml 10%
Solution, room temperature mechanical stir 6 h, after the completion of reaction, are washed with water；
（5）The material of synthesis is added in 30 ml distilled water, adds in 0.02 g of laccase powder, it is 120 to be then placed in rotating speed
In the shaking table of rpm, 30 DEG C of 6 h of reaction；
（6）Finally with PH=7.0, three times, removal fails to combine resolvase up, receives for the phosphate buffer washing of 0.1 mol/l
Collect immobilization laccase, it is spare.
The carboxyl carbon nanotube coated with polymethyl methacrylate of preparation is placed in scanning electron microscope（SEM）Under
The morphology and grain size of material are observed, finds to be attached with polymethyl methacrylate in carboxyl carbon nanotube, shows poly- methyl-prop
E pioic acid methyl ester is aggregated in carboxyl carbon nanotube, and shows cavernous structure, as shown in Figure 1.
Embodiment 2：
（1）It weighs the above-mentioned carboxyl carbon nanotubes obtained by the reaction of 0.2 g to be distributed in the normal octane solution of 5 mL 6%, adds in 0.2
G lauryl sodium sulfate and 30 mL water, molten ultrasonic 15 min of ice；
（2）4 mL methyl methacrylates, 1 mL divinylbenzenes, 8 mL1% potassium peroxydisulfates are added in, 70 DEG C of reaction temperature is stirred
24 h of polymerization are mixed, after the completion of reaction, are naturally cooling to room temperature, are centrifuged；
（3）The obtained solid water and ethyl alcohol of above-mentioned reaction alternately wash, and are dried in vacuo, cooling, collect product and obtain black
Solid particle is the carboxyl carbon nanotube coated with polymethyl methacrylate；
（4）The carboxyl carbon nanotube that 0.2 g polymethyl methacrylates coat is weighed, adds in the glutaraldehyde water of 20 ml 10%
Solution, room temperature mechanical stir 6 h, after the completion of reaction, are washed with water；
（5）The material of synthesis is added in 30 ml distilled water, adds in 0.02 g of laccase powder, it is 120 to be then placed in rotating speed
In the shaking table of rpm, 30 DEG C of 6 h of reaction；
（6）Finally with PH=7.0, three times, removal fails to combine resolvase up, receives for the phosphate buffer washing of 0.1 mol/l
Collect immobilization laccase, it is spare.
In order to further determine polymethyl methacrylate whether coated carboxyl carbon nanotube, by prepared sample into
Row X-ray diffractogram spectrum analysis.As shown in Fig. 2, compared with carboxyl carbon nanotube, with the carboxylic of polymethyl methacrylate cladding
Base carbon nanotube has lacked 25.9 ° of peak, this shows polymethyl methacrylate coated carboxyl carbon nanotube.
Embodiment 3：
（1）It weighs the above-mentioned carboxyl carbon nanotubes obtained by the reaction of 0.2 g to be distributed in the normal octane solution of 5 mL 6%, adds in 0.2
G lauryl sodium sulfate and 30 mL water, molten ultrasonic 15 min of ice；
（2）4 mL methyl methacrylates, 1 mL divinylbenzenes, 8 mL1% potassium peroxydisulfates are added in, 70 DEG C of reaction temperature is stirred
24 h of polymerization are mixed, after the completion of reaction, are naturally cooling to room temperature, are centrifuged；
（3）The obtained solid water and ethyl alcohol of above-mentioned reaction alternately wash, and are dried in vacuo, cooling, collect product and obtain black
Solid particle is the carboxyl carbon nanotube coated with polymethyl methacrylate；
（4）The carboxyl carbon nanotube that 0.2 g polymethyl methacrylates coat is weighed, adds in the glutaraldehyde water of 20 ml 10%
Solution, room temperature mechanical stir 6 h, after the completion of reaction, are washed with water；
（5）The material of synthesis is added in 30 ml distilled water, adds in 0.02 g of laccase powder, it is 120 to be then placed in rotating speed
In the shaking table of rpm, 30 DEG C of 6 h of reaction；
（6）Finally with PH=7.0, three times, removal fails to combine resolvase up, receives for the phosphate buffer washing of 0.1 mol/l
Collect immobilization laccase, it is spare.
In order to which verify cladding carboxyl carbon nanotube is polymethyl methacrylate, it is red that prepared sample is subjected to Fourier
External spectrum is analyzed.As shown in figure 3, compared with carboxyl carbon nanotube, with the carboxyl carbon nanometer of polymethyl methacrylate cladding
The peak of pipe changes, in 2900 cm-1Observe the feature of carboxyl carbon nanotube coated with polymethyl methacrylate in place
Band corresponds to methylmethylene.In addition, it is 1700 cm in wavelength -1, 1200 cm-1Place is respectively present absorption peak.This shows to gather
Methyl methacrylate coated carboxyl carbon nanotube.
Embodiment 4：
（1）It weighs the above-mentioned carboxyl carbon nanotubes obtained by the reaction of 0.2 g to be distributed in the normal octane solution of 5 mL 6%, adds in 0.2
G lauryl sodium sulfate and 30 mL water, molten ultrasonic 15 min of ice；
（2）4 mL methyl methacrylates, 1 mL divinylbenzenes, 8 mL1% potassium peroxydisulfates are added in, 70 DEG C of reaction temperature is stirred
24 h of polymerization are mixed, after the completion of reaction, are naturally cooling to room temperature, are centrifuged；
（3）The obtained solid water and ethyl alcohol of above-mentioned reaction alternately wash, and are dried in vacuo, cooling, collect product and obtain black
Solid particle is the carboxyl carbon nanotube coated with polymethyl methacrylate；
（4）The carboxyl carbon nanotube that 0.2 g polymethyl methacrylates coat is weighed, adds in the glutaraldehyde water of 20 ml 10%
Solution, room temperature mechanical stir 6 h, after the completion of reaction, are washed with water；
（5）The material of synthesis is added in 30 ml distilled water, adds in 0.02 g of laccase powder, it is 120 to be then placed in rotating speed
In the shaking table of rpm, 30 DEG C of 6 h of reaction；
（6）Finally with PH=7.0, three times, removal fails to combine resolvase up, receives for the phosphate buffer washing of 0.1 mol/l
Collect immobilization laccase, it is spare.
（7）0.05 g immobilization laccases are weighed, is put into II solution of gold orange of a concentration of 10 mg/L of 20 mL, is in rotating speed
120 r/min, temperature are 30 DEG C, and 24 h are reacted in pH=7.Then it collects supernatant and surveys absorbance.
In order to study decolorizing effect of the immobilization laccase to gold orange II, immobilization laccase is used for decolorization experiment.Such as Fig. 4
（a）It is shown, it is 30 DEG C in temperature, the methylenum careuleum percent of decolourization of immobilization laccase is 73.72%.Such as Fig. 4（b）It is shown, fixing laccase
36.89% is higher than to the percent of decolourization of gold orange II after recycling 10 times.
Claims (8)
1. application of a kind of immobilization laccase in gold orange II is decolourized, it is characterised in that this method includes the following steps：
(1) the above-mentioned carboxyl carbon nanotubes obtained by the reaction of 0.2g are weighed to be distributed in the normal octane solution of 5mL 6%, add in 0.2g
Lauryl sodium sulfate and 30mL water, the molten ultrasound 15min of ice；
(2) 4mL methyl methacrylates, 1mL divinylbenzenes, 8mL1% potassium peroxydisulfates, 70 DEG C of reaction temperature are added in, stirring gathers
It closes for 24 hours, after the completion of reaction, is naturally cooling to room temperature, centrifuge；
(3) the obtained solid water of above-mentioned reaction and ethyl alcohol alternately wash, and are dried in vacuo, cooling, collect product and obtain black
Solid particle is the carboxyl carbon nanotube coated with polymethyl methacrylate；
(4) the carboxyl carbon nanotube that 0.2g polymethyl methacrylates coat is weighed, the glutaraldehyde for adding in 20ml 10% is water-soluble
Liquid, room temperature mechanical stirring 6h, after the completion of reaction, is washed with water；
(5) material of synthesis is added in 30ml distilled water, adds in laccase powder 0.02g, be then placed in rotating speed as 120rpm
Shaking table in, 30 DEG C reaction 6h；
(6) finally with PH=7.0, three times, removal fails to combine resolvase up, receives for the phosphate buffer washing of 0.1mol/l
Collect immobilization laccase, it is spare.
(7) 0.05g immobilization laccases are weighed, are put into II solution of gold orange of a concentration of 10mg/L of 20mL, are 120r/ in rotating speed
Min, temperature are 30 DEG C, pH=7, and reaction is for 24 hours.Then it collects supernatant and surveys absorbance.
2. preparation method according to claim 1, which is characterized in that the ultrasound described in step (1) is received to carboxyl carbon
First then ultrasound 2min, the dispersibility of guarantee carboxyl carbon nanotube in the solution add when mitron is distributed to normal octane solution
Other reagents carry out the molten ultrasound 15min of ice.
3. carboxyl carbon nanotube used in step (1) has larger specific surface area, polymethyl methacrylate can be filled
In the dispersin polymerization to carboxyl carbon nanotube divided, the surface area of carrier is improved.
4. preparation method according to claim 1, which is characterized in that prepared carboxyl carbon nanotube exists in step (1)
A concentration of 40mg/ml of dispersion liquid in 6% normal octane.
5. preparation method according to claim 1, which is characterized in that polymeric reaction temperature should be in always in step (2)
At 70 DEG C.
6. preparation method according to claim 1, which is characterized in that in the middle water of step (3) and ethanol wash process, need
6h is fully first impregnated with ethyl alcohol, ensures fully removal impurity.
7. preparation method according to claim 1, which is characterized in that polymethyl methacrylate and laccase in step (5)
Mass ratio be 10：1.
8. preparation method according to claim 1, which is characterized in that it is terrible to survey supernatant absorbance in step (7) to be
Go out the concentration of II solution of the gold orange after decolorization, and then calculate percent of decolourization.
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| CN201711390178.1A CN108217972A (en) | 2017-12-21 | 2017-12-21 | A kind of application of immobilization laccase in gold orange II is decolourized |
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| CN106434621A (en) * | 2016-10-28 | 2017-02-22 | 华北电力大学 | Method for immobilizing laccase on polyacrylamide-coated magnetic nano particles and application of method |
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2017
- 2017-12-21 CN CN201711390178.1A patent/CN108217972A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1640933A (en) * | 2004-01-13 | 2005-07-20 | 中国科学院成都有机化学有限公司 | Method for preparing polymer/carbon nano composite material |
| WO2007001355A2 (en) * | 2004-09-08 | 2007-01-04 | Rensselaer Polytechnic Institute | Enhanced stability of proteins immobilized on nanoparticles |
| CN102260662A (en) * | 2010-05-24 | 2011-11-30 | 中国科学院过程工程研究所 | Carrier used for immobilized enzymes, its purpose and carrier fixed with enzymes |
| CN102373192A (en) * | 2011-01-21 | 2012-03-14 | 华东理工大学 | Immobilization method of proteinase molecule by using nano-material and application thereof |
| CN106434621A (en) * | 2016-10-28 | 2017-02-22 | 华北电力大学 | Method for immobilizing laccase on polyacrylamide-coated magnetic nano particles and application of method |
Non-Patent Citations (2)
| Title |
|---|
| 曾涵 等: "《高分子聚合物-多壁碳纳米管复合物固定漆酶及其在玻碳电极上的直接电子转移》", 《应用化学》 * |
| 陈坚 等: "《环境生物技术应用与发展》", 30 June 2001, 中国轻工业出版社 * |
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